Conclusiones

1.3.1. Pantothenic Acid. Pantothenic acid also called as pantothenate or vitamin B5 is a water soluble dihydroxy carboxylic acid with an internal amide bond that links D-pantoate and β-alanine residues (Figure 1.12a). It is commercially available is as the calcium salt (Figure 1.12b) [54].

NH OH

Figure 1.12: Molecular structure of (a) Pantothenic acid [C₉H₁₇NO₅] and (b) Pantothenic acid calcium salt [C18H32CaN2O10].

1.3.2. Biological Role of Pantothenic Acid. PA is a component of Coenzyme-A (CoA) and also it is required for its biosynthesis [57]. As a result pantothenic acid participates in many metabolic processes [55]. It participates in synthesis of numerous enzymes and helps maintain precise communication between the central nervous system and the brain. Only the dextrorotatory (D) isomer of PA possesses biologic activity. The levorotatory (L) form may antagonize the effects of the dextrorotatory isomer [56].

1.3.3 Coenzyme A. CoA is a coenzyme participates in the synthesis and oxidation of fatty acids. CoA acts as a carrier of acyl group to form acetyl-CoA, a way to transport carbon atoms within the cell [54]. It assists in transferring fatty acids from the cytoplasm to mitochondria. CoA plays a significant role in metabolism of steroids, fatty acids and phosphatides and also in synthesis of carbohydrates, fats and proteins [58]. PA in the form of CoA (Figure 1.13) is required for acylation and acetylation which are involved in signal transduction and enzyme activation and deactivation, respectively. PA is liberated from CoA by using various enzymes like Pyrophosphatase, Ortho-Phosphatase and Pantetheinase [59, 60].

Figure 1.13: Molecular structure of Coenzyme A.

1.3.4. Deficiency and Toxicity of Pantothenic Acid. PA is an essential nutrient found at low concentrations in all animals and plant tissues. The common sources of PA are milk, corn, eggs, meats, peanuts, peas, soybeans, salmon and wheat germ. Because of its wide occurrence, the deficiency of PA is very rare unless specifically engineered for the purposes of biochemical investigations. A deficiency of PA in adults is virtually nonexistent, largely because it is present in so many foods. PA deficiency reduces biologic actylations and leads to variety of pathologic changes in blood cholesterol partition and antibody production. PA deficiency particularly affects nervous system and the adrenal cortex. It also causes fatigue, heart problems, increased risk of infections, abdominal pains, sleep disturbances, numbness and altered sensation in the arms and legs, muscle weakness, cramps, increased sensitivity to insulin (the hormone that lowers blood glucose levels), decreased blood cholesterol levels and decreased potassium levels in the body [61-63]. The normal daily allowance for PA in adults is about 4 to 7 mg per day.

PA are non toxic, no serious side effects have been reported, even at intakes of up to 10 g per day. Very large amounts of PA (>10g per day) can cause diarrhea and no other adverse reactions to high doses have been reported.

Pantothenic Acid

1.3.5. Pantothenic Acid as Animal Feed Supplement. Since PA is essential in metabolism due to its incorporation into CoA and acyl-carrier-protein it is supplemented in animal feed particularly for dairy cows. Dairy cows have two sources of PA: on the one hand fodder, and on the other hand, ruminal PA synthesis by microorganisms [64]. The ruminal synthesis of PA does not cover the proposed daily PA requirements for tissue and for milk production. Because PA is ubiquitous in feedstuffs, requirements may be met by PA intake via feed provided that feed-bound PA is not degraded in the rumen. Small intestine is the major site of PA absorption in ruminants [65]. Synthesis of PA in animals appears to be influenced by forage to concentrate ratio in the diet. It is not yet clear, if oral PA supplementations can increase the duodenal PA flow in dairy cows, but it has been reported that about 80% of supplemented PA disappears between the mouth and duodenum. Amount of PA can be a measured in blood to investigate whether PA intake via feed might meet the proposed requirements. However, supplementation of PA can increase blood PA levels [66].

Currently there are no efficient and sensitive methods available for the quantification of PA in complex matrix like bovine serum. Therefore, the objective of the present work was to develop a quantitative and sensitive method for the determination and quantification of PA in bovine serum.

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PAPER

I. DEVELOPMET AD EVALUATIO OF A EFFICIET PROCESS FOR THE